| A novel micropit device integrates automated cell positioning by dielectrophoresis and nuclear transfer by electrofusion. | |
| | |
MedLine Citation:
|
PMID: 20499188 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Nuclear transfer (NT) cloning involves manual positioning of individual donor-recipient cell couplets for electrofusion. This is time-consuming and introduces operator-dependent variation as a confounding parameter in cloning trials. In order to automate the NT procedure, we developed a micro-fluidic device that integrates automated cell positioning and electrofusion of isolated cell couplets. A simple two layer micro-fluidic device was fabricated. Thin film interdigitated titanium electrodes (300 nm thick, 250 microm wide and 250 microm apart) were deposited on a solid borosilicate glass substrate. They were coated with a film of electrically insulating photosensitive epoxy polymer (SU-8) of either 4 or 22 microm thickness. Circular holes ("micropits") measuring 10, 20, 30, 40 or 80 microm in diameter were fabricated above the electrodes. The device was immersed in hypo-osmolar fusion buffer and manually loaded with somatic donor cells and recipient oocytes. Dielectrophoresis (DEP) was used to attract cells towards the micropit and form couplets on the same side of the insulating film. Fusion pulses between 80 V and 120 V were applied to each couplet and fusion scored under a stereomicroscope. Automated couplet formation between oocytes and somatic cells was achieved using DEP. Bovine oocyte-oocyte, oocyte-follicular cells and oocyte-fibroblast couplets fused with up to 69% (n = 13), 50% (n = 30) and 78% (n = 9) efficiency, respectively. Fusion rates were comparable to parallel plate or film electrodes that are conventionally used for bovine NT. This demonstrates proof-of-principle that a micropit device is capable of both rapid cell positioning and fusion. |
| | |
Authors:
|
Andrew L Clow; Paul T Gaynor; Björn J Oback |
Related Documents
:
|
1701988 - Characterization of cell fusion in xc cells induced by suncus murinus mammary tumor virus. 1473148 - Lysosome recruitment and fusion are early events required for trypanosome invasion of m... 18031758 - Functional roles of the hook in a rotating tethered cell. 1901498 - Small unilamellar vesicles are able to fuse with mycoplasma capricolum cells. 23457478 - Phosphorescent imaging of living cells using a cyclometalated iridium(iii) complex. 21853128 - Mitochondrial dna mutations regulate metastasis of human breast cancer cells. |
Publication Detail:
|
Type: Journal Article |
Journal Detail:
|
Title: Biomedical microdevices Volume: 12 ISSN: 1572-8781 ISO Abbreviation: Biomed Microdevices Publication Date: 2010 Oct |
Date Detail:
|
Created Date: 2010-08-05 Completed Date: 2011-01-10 Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 100887374 Medline TA: Biomed Microdevices Country: United States |
Other Details:
|
Languages: eng Pagination: 777-86 Citation Subset: IM |
Affiliation:
|
Department of Electrical and Computer Engineering, University of Canterbury, Private Bag 4800, Christchurch 8140, New Zealand. alc76@student.canterbury.ac.nz |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Automation Cattle Cell Fusion / instrumentation* Electrodes Electrophoresis / instrumentation* Female Microfluidic Analytical Techniques* Models, Theoretical Nuclear Transfer Techniques / instrumentation* Oocysts / cytology Systems Integration |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Designing Rites to Re-enchant Secularized Society: New Varieties of Spiritualized Therapy in Contemp...
Next Document: Molecular characterization and virulence of Beauveria spp. from the pine processionary moth, Thaumet...